Novel sericin-based hepatocyte serum-free medium and sericin’s effect on hepatocyte transcriptome

Table 1 Medium formula of each group in part 1

Group A	Group B	Group C	Group D
DMEM/F12	HepatoZYME	DMEM/F12	DMEM/F12
ITS 10 mL/L	L-Glutamine 2 μmol/mL	FBS 100 mL/L	PEN-SM 100 U/mL
HGF 20 ng/mL	PEN-SM 100 U/mL	PEN-SM 100 U/mL
EGF 10 ng/mL
Dex 1 nmol/mL
Sericin 2 mg/mL
PEN-SM 100 U/mL

ITS: Insulin-transferrin-sodium; HGF: Hepatocyte growth factor; EGF: Epidermal growth factor; Dex: Dexamethasone; PEN-SM: Penicillin-streptomycin; FBS: Fetal bovine serum.

Table 2 Medium formula of each group in part 2

Group A (sericin)	Group B (control)
DMEM/F12	DMEM/F12
ITS 10 mL/L	ITS 10 mL/L
HGF 20 ng/mL	HGF 20 ng/mL
EGF 10 ng/mL	EGF 10 ng/mL
Dex 1 nmol/mL	Dex 1 nmol/mL
Sericin 2 mg/mL	PEN-SM 100 U/mL
PEN-SM 100 U/mL

ITS: Insulin-transferrin-sodium; HGF: Hepatocyte growth factor; EGF: Epidermal growth factor; Dex: Dexamethasone; PEN-SM: Penicillin-streptomycin; FBS: Fetal bovine serum.

Table 3 Up-regulated genes associated with cell proliferation

Up-regulated genes	Pathways involved	Fold change
CCR6	Cytokine-cytokine receptor interaction	1.99
EGFR	Pathways in cancer, MAPK signaling pathway	1.55
FOS	Pathways in cancer, MAPK signaling pathway	1.59

Table 4 Comparison of CCR6 expression and genes in the MAPK pathway

Genes	FC in gene chip	FC1	FC2	FC3	FC4
CCR6	1.99	1.36 ± 0.091	1.40 ± 0.01	2.36 ± 0.041	0.75 ± 0.01
EGFR	1.55	1.76 ± 0.041	7.71 ± 0.491	1.46 ± 0.131	2.34 ± 0.241
FOS	1.59	1.22 ± 0.091	0.96 ± 0.07	1.77 ± 0.051	0.96 ± 0.01
Src	1.15	0.84 ± 0.161	2.44 ± 0.081	1.93 ± 0.701	3.67 ± 0.401
PI3K	1.24	3.00 ± 0.271	3.01 ± 0.391	1.43 ± 0.15	3.10 ± 0.141
AKT1	1.13	1.22 ± 0.041	1.21 ± 0.051	2.04 ± 0.101	1.59 ± 0.061
JNK1	1.09	0.98 ± 0.07	1.07 ± 0.11	2.69 ± 0.091	2.09 ± 0.041
NFκB1	1.03	0.97 ± 0.07	1.51 ± 0.021	1.77 ± 0.041	0.84 ± 0.02
MMP-9	1.22	1.38 ± 0.151	1.10 ± 0.06	1.51 ± 0.041	1.56 ± 0.041
GRB2	1.05	1.04 ± 0.10	1.48 ± 0.031	1.34 ± 0.011	1.06 ± 0.05
SHC2	1.30	0.98 ± 0.03	1.13 ± 0.06	1.39 ± 0.211	1.47 ± 0.171
K-RAS	1.09	1.16 ± 0.05	1.21 ± 0.081	1.42 ± 0.041	1.53 ± 0.011
RAF1	1.01	1.09 ± 0.10	1.24 ± 0.021	1.48 ± 0.021	1.22 ± 0.031
MEK2	1.02	5.86 ± 1.221	4.61 ± 1.111	2.38 ± 1.051	8.45 ± 1.001
ERK1	1.07	1.80 ± 0.621	1.29 ± 0.031	1.02 ± 0.26	1.19 ± 0.02
ERK2	1.18	2.58 ± 0.211	2.52 ± 0.621	5.15 ± 0.261	1.63 ± 0.061
c-myc	1.17	2.32 ± 0.211	1.67 ± 0.271	1.41 ± 0.301	0.06 ± 0.011
cyclinE1	1.04	1.04 ± 0.13	1.52 ± 0.031	2.21 ± 0.021	1.48 ± 0.011

¹Significant difference between group A and group B at the same incubation time (P < 0.05). FC1, FC2, FC3, FC4 was the fold change (group A/group B) of gene expression measured by RT-qPCR on days 1, 2, 3, 4, respectively. Data are expressed as mean ± SD (n = 3).